自噬对大鼠肾缺血再灌注损伤的保护作用及机制

发布时间：2018-09-11 10:25

【摘要】：目的观察大鼠肾脏缺血再灌注(ischemia reperfusion,I/R)损伤中肾小管上皮细胞的自噬激活情况,探讨其对肾脏I/R损伤发挥保护性作用的分子机制。方法将40只雄性Wistar大鼠随机分为4组:假手术组(Sham)、I/R组、氯喹干预组(I/R+CQ)和雷帕霉素干预组(I/R+Rap)。常规方法建立大鼠肾脏I/R损伤模型,再灌注24 h后留取各组大鼠血和肾脏标本。血标本用于尿素氮(BUN)和血肌酐(Scre)含量检测;肾组织标本行HE染色,观察病理学损伤情况;TUNEL法检测肾小管上皮细胞凋亡的改变;采用免疫组化法检测Beclin-1和Caspase-3蛋白表达,透射电镜观察大鼠肾脏自噬泡的形成情况。结果与I/R组相比,I/R+CQ组BUN和Scre明显升高(P0.05,P0.01),肾组织损伤积分增加(P0.01),TUNEL阳性细胞数增多(P0.05),Beclin-1蛋白表达减弱(P0.01),Caspase-3蛋白表达增强(P0.01),电镜下自噬泡数量较少(P0.05);I/R+Rap组BUN和Scre显著降低(P0.01),肾组织损伤积分减低(P0.05),TUNEL阳性细胞数减少(P0.05),Beclin-1蛋白表达增强(P0.01),Caspase-3蛋白表达减弱(P0.01),自噬泡数量明显增多(P0.01)。结论自噬激活在肾脏I/R损伤过程中可通过抑制凋亡而发挥的保护作用,其机制可能涉及Beclin-1及Caspase-3等蛋白的表达调控。
[Abstract]:Objective to observe the activation of autophagy of renal tubular epithelial cells during renal ischemia-reperfusion (ischemia reperfusion,I/R) injury in rats and to explore the molecular mechanism of its protective effect on renal I / R injury. Methods Forty male Wistar rats were randomly divided into four groups: sham operation group (Sham) / I / R group), chloroquine intervention group (I / R CQ) and rapamycin intervention group (I / R Rap). The rat kidney I / R injury model was established by routine method. The blood and kidney samples of each group were collected after 24 h reperfusion. Blood samples were used to detect the contents of urea nitrogen (BUN) and creatinine (Scre). Renal tissue specimens were stained with HE to observe the pathological damage and Tunel method was used to detect the apoptosis of renal tubular epithelial cells. The expression of Beclin-1 and Caspase-3 protein was detected by immunohistochemical method. The formation of autophagy in rat kidney was observed by transmission electron microscope. Results compared with the I / R group, the BUN and Scre in the I / R CQ group were significantly higher (P0.05, P0.01), the number of Tunel positive cells was increased (P0.01) (P0.05), the expression of Caspase-3 protein was significantly decreased (P0.01), and the number of autophagic vesicles was significantly decreased in the IP-R Rap group (P0.05). (P0.01), the number of Tunel positive cells decreased (P0.05), the expression of Beclin-1 protein decreased (P0.01) and the number of autophagy increased significantly (P0.01). Conclusion autophagy may play a protective role in the process of renal I / R injury by inhibiting apoptosis, and its mechanism may be related to the regulation of Beclin-1 and Caspase-3 protein expression.
【作者单位】：北京大学第三医院检验科;
【基金】：国家自然科学基金(81271903) 国家科技支撑计划课题(2012BA137B01)
【分类号】：R692

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